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The honey bee queen is the central hub of a colony to produce eggs and release pheromones to maintain social cohesion. Among many environmental stresses, viruses are a major concern to compromise the queen’s health and reproductive vigor. Viruses have evolved numerous strategies to infect queens either via vertical transmission from the queens’ parents or horizontally through the worker and drones with which she is in contact during development, while mating, and in the reproductive period in the colony. Over 30 viruses have been discovered from honey bees but only few studies exist on the pathogenicity and direct impact of viruses on the queen’s phenotype. An apparent lack of virus symptoms and practical problems are partly to blame for the lack of studies, and we hope to stimulate new research and methodological approaches. To illustrate the problems, we describe a study on sublethal effects of Israeli Acute Paralysis Virus (IAPV) that led to inconclusive results. We conclude by discussing the most crucial methodological considerations and novel approaches for studying the interactions between honey bee viruses and their interactions with queen health.

In the case of SARS-CoV-2 pandemic management, wastewater-based epidemiology aims to derive information on the infection dynamics by monitoring virus concentrations in the wastewater. However, due to the intrinsic random fluctuations of the viral signal in wastewater caused by several influencing factors that cannot be determined in detail (e.g. dilutions; number of people discharging; variations in virus excretion; water consumption per day; transport and fate processes in sewer system), the subsequent prevalence analysis may result in misleading conclusions. It is thus helpful to apply data filtering techniques to reduce the noise in the signal. In this paper we investigate 13 smoothing algorithms applied to the virus signals monitored in four wastewater treatment plants in Austria. The parameters of the algorithms have been defined by an optimization procedure aiming for performance metrics. The results are further investigated by means of a cluster analysis. While all algorithms are in principle applicable, SPLINE, Generalized Additive Model and Friedman's Super Smoother are recognized as superior methods in this context (with the latter two having a tendency to over-smoothing). A first analysis of the resulting datasets indicates the positive effect of filtering to the correlation of the viral signal to monitored incidence values.

A simple model is designed for an inductive immunosensor in which the magnetic particles are attached to the bioreceptors to form a sandwich on the surface of an inductor. The inductor consists of a coil covered on a silicon oxide wafer. The coil comprises 250 turns of a planar gold wire, which is approximately 200 nm thick and 392 mm long, placed in a circle with a diameter of 2 mm. The model is well characterised by controlling the geometrical and electrical parameters and also the permeability of the magnetic material. To evaluate the feasibility of the model for virus monitoring, a novel inductive immunosensor is designed and for the first time applied for the detection of hepatitis B surface antigen (HBsAg). At first, Fab′ segment of primary anti-HBsAg is immobilised on the coil. Then, the coil is exposed to HBsAg and the complex is introduced to a secondary antibody conjugated with magnetic particles to form an immune-sandwich. Finally, the influence of magnetic particles on the coil inductance is recorded and used as a signal for HBsAg detection. The magnetic inductive immunosensor showed specific responses toward HBsAg with the detection limit of 1 ng mL −1 , linear range of 1 to 200 ng mL −1 , and a sensitivity of 6 × 10 −4  mL ng −1 . The experimental results showed a very good agreement with simulation data indicating the compatibility of sensor sensitivity to the expected theoretical values. Graphical abstract

Oncolytic adenoviruses are in development as immunotherapeutic agents for solid tumors. Their efficacy is in part dependent on their ability to replicate in tumors. It is, however, difficult to obtain evidence for intratumoral oncolytic adenovirus replication if direct access to the tumor is not possible. Detection of systemic adenovirus DNA, which is sometimes used as a proxy, has limited value because it does not distinguish between the product of intratumoral replication and injected virus that did not replicate. Therefore, we investigated if detection of virus-associated RNA (VA RNA) by RT-qPCR on liquid biopsies could be used as an alternative. We found that VA RNA is expressed in adenovirus-infected cells in a replication-dependent manner and is secreted by these cells in association with extracellular vesicles. This allowed VA RNA detection in the peripheral blood of a preclinical in vivo model carrying adenovirus-injected human tumors and on liquid biopsies from a human clinical trial. Our results confirm that VA RNA detection in liquid biopsies can be used for minimally invasive assessment of oncolytic adenovirus replication in solid tumors in vivo.

The COVID-19 pandemic remains a constant threat to human health, the economy, and social relations. Scientists around the world are constantly looking for new technological tools to deal with the pandemic. Such tools are the rapid virus detection tests, which are constantly evolving and optimizing. This paper presents a biosensor platform for the rapid detection of spike protein both in laboratory conditions and in swab samples from hospitalized patients. It is a continuation and improvement of our previous work and consists of a microcontroller-based readout circuit, which measures the capacitance change generated in an interdigitated electrode transducer by the presence either of sole spike protein or the presence of SARS-CoV-2 particles in swab samples. The circuit efficiency is calibrated by its correlation with the capacitance measurement of an LCR (inductance (L), capacitance (C), and resistance (R)) meter. The test result is made available in less than 2 min through the microcontroller’s LCD (liquid-crystal display) screen, whereas at the same time, the collected data are sent wirelessly to a mobile application interface. The novelty of this research lies in the potential it offers for continuous and effective screening of SARS-CoV-2 patients, which is facilitated and enhanced, providing big data statistics of COVID-19 in terms of space and time. This device can be used by individuals for SARS-CoV-2 testing at home, by health professionals for patient monitoring, and by public health agencies for monitoring the spatio-temporal spread of the virus.

Background: Hepatitis C virus (HCV) represents a serious worldwide healthcare problem. No protective vaccines against HCV have been developed yet due to the fact that HCV is rapidly mutable, allowing the virus to escape from the neutralizing antibodies. Understanding of HCV was initially hampered by the inability to achieve viral replication in cell culture. Given its essential roles in viral polyprotein processing and immune evasion, HCV NS3/4A protease is a prime target for antiviral chemotherapy. We aimed to establish in vivo cell-based assay system for monitoring the activity of NS3/4A protease from HCV genotype 4a, the predominant genotype in Egypt, and the Middle East. Furthermore, the developed system was used to evaluate the inhibitory potency of a series of computer-designed chemically-synthesized compounds against NS3/4A protease from HCV genotype 4a. Materials and Methods: Native as well as mutant cleavage sites to NS3/4A protease were cloned in frame into β-galactosidase gene of TA cloning vector. The target specificity of HCV NS3/4A was evaluated by coexpression of β-galactosidase containing the protease cleavage site with NS3/4A protease construct in bacterial cells. The activity of β-galactosidase was colorimetrically estimated in the cell lysate using orthonitro phenyl β-D-galactopyanoside (ONPG) as a substrate. Results and Conclusions: We successfully developed an efficient cell-based system based on the blue/white selection of bacterial cells that are able to express functional/nonfunctional β-galactosidase enzyme.

Abstract The disease most dangerous for the cultivation activity is virus. Viruses are organisms subseluler that contain only nucleic acid (RNA or DNA) as genetic material. Koi Herpes Virus is one type of virus that causes mortality in cultured Cyprinids. KHV disease in Indonesia started in Blitar, East Java on March 2002 because the entry of imported koi fish that carry the virus KHV, while mortality prosentase could reach 80% - 85%, which causes loss of about 5 billion rupiah. In addition of KHV, there are several types of viral diseases in shrimp is White Spot Syndrome Virus (WSSV), Taura Syndrome Virus (TSV), dan Yellow Head Virus (YHV). Disease can cause losses in farming activities, such as WSSV. WSSV is an endemic disease since 1995. disease WSSV is exotic viral disease that attacks the shrimp monodon in 1998/1999 has resulted in decreased production of very large, so the Indonesian shrimp exports down 33,000 tons. Treatment of viral diseases is difficult because the virus resistant to certain antibiotics and chemical compounds. Therefore, prevention needs to be done, one through the monitoring activities conducted on the northern coast of East Java. The method implemented is monitoring in location and identification of viruses by PCR (Polymerase Chain Reaction). Monitoring in location includes water quality measurements and sampling. Identification of virus carried by IQ 2000TM. The identification procedure includes extraction, amplification and electrophoresis. Regional monitoring conducted on the northern coast of East Java includes Gresik, Lamongan, Tuban, Bangkalan, Sampang, Pamekasan, and Sumenep. Water quality at locations quite well. Results activities of monitoring on the northern coast of East Java is disease White Spot Syndrome Virus (WSSV) was found positive in several locations: Gresik, Lamongan and Tuban, while the virus Taura Syndrome Virus (TSV) and Yellow Head Virus (YHV) was not found at all locations . In tilapia, disease Koi Herpes Virus (KHV) was found positive in Tuban.

H7N9 avian influenza has broken out in Chinese poultry 10 times since 2013 and impacted the industry severely. Although the epidemic is currently under control, there is still a latent threat. Epidemiological surveillance data for non-human H7N9 avian influenza from April 2013 to April 2020 were used to analyse the regional distribution and spatial correlations of positivity rates in different months and years and before and after comprehensive immunisation. In addition, positivity rate monitoring data were disaggregated into a low-frequency and a high-frequency trend sequence by wavelet packet decomposition (WPD). The particle swarm optimisation algorithm was adopted to optimise the least squares support-vector machine (LS-SVM) model parameters to predict the low-frequency trend sequence, and the autoregressive integrated moving average (ARIMA) model was used to predict the high-frequency one. Ultimately, an LS-SVM-ARIMA combined model based on WPD was constructed. The virus positivity rate was the highest in late spring and early summer, and overall it fell significantly after comprehensive immunisation. Except for the year 2015 and the single month of December from 2013 to 2020, there was no significant spatiotemporal clustering in cumulative non-human H7N9 avian influenza virus detections. Compared with the ARIMA and LS-SVM models, the LS-SVM-ARIMA combined model based on WPD had the highest prediction accuracy. The mean absolute and root mean square errors were 2.4% and 2.0%, respectively. Low error measures prove the validity of this new prediction method and the combined model could be used for inference of future H7N9 avian influenza virus cases. Live poultry markets should be closed in late spring and early summer, and comprehensive H7N9 immunisation continued.

The COVID-19 pandemic has had a profound impact on various aspects of our lives, affecting personal, occupational, economic, and social spheres. Much has been learned since the early 2020s, which will be very useful when the next pandemic emerges. In general, mobility and virus spread are strongly related. However, most studies analyze the impact of COVID-19 on mobility, but not much research has focused on analyzing the impact of mobility on virus transmission, especially from the point of view of monitoring virus incidence, which is extremely important for making sound decisions to control any epidemiological threat to public health. As a result of a thorough analysis of COVID-19 and mobility data, this work introduces a novel measure, the Infection Ratio (IR), which is not sensitive to underestimation of positive cases and is very effective in monitoring the pandemic’s upward or downward evolution when it appears to be more stable, thus anticipating possible risk situations. For a bounded spatial context, we can infer that there is a significant threshold in the restriction of mobility that determines a change of trend in the number of infections that, if maintained for a minimum period, would notably increase the chances of keeping the spread of disease under control. Results show that IR is a reliable indicator of the intensity of infection, and an effective measure for early monitoring and decision making in smart cities.

In microbiological research, it is important to understand the time course of each step in a pathogen’s lifecycle and changes in the host cell environment induced by infection. This study is the first to develop a real-time monitoring system that kinetically detects luminescence reporter activity over time without sampling cells or culture supernatants for analyzing the virus replication. Subgenomic replicon experiments with hepatitis C virus (HCV) showed that transient translation and genome replication can be detected separately, with the first peak of translation observed at 3–4 h and replication beginning around 20 h after viral RNA introduction into cells. From the bioluminescence data set measured every 30 min (48 measurements per day), the initial rates of translation and replication were calculated, and their capacity levels were expressed as the sums of the measured signals in each process, which correspond to the areas on the kinetics graphs. The comparison of various HuH-7-derived cell lines showed that the bioluminescence profile differs among cell lines, suggesting that both translation and replication capacities potentially influence differences in HCV susceptibility. The effects of RNA mutations within the 5′ UTR of the replicon on viral translation and replication were further analyzed in the system developed, confirming that mutations to the miR-122 binding sites primarily reduce replication activity rather than translation. The newly developed real-time monitoring system should be applied to the studies of various viruses and contribute to the analysis of transitions and progression of each process of their life cycle.